Members of the bacterial genus Vibrio have long been recognized as the causative agent of a variety of gastrointestinal disorders associated with ingestion of contaminated seafoods. It is the goal of this project to develop a highly sensitive enzyme immunoassay with an amplified indicator system for the detection of vibrios in seafood products and marine waters. Vibrio parahaemolyticus will be used as the model organism. Antibodies specific for the model organism will be produced and conjugates of these antibodies will be prepared for use in the enzyme cycling indicator systems to be used. The assay protocol will be developed in a vacuum microwell format, using Vibrio parahaemolyticus from pure cultures as antigen, and will be evaluated for sensitivity and specificity. Various methods of rapidly extracting and concentrating vibrios from samples will be compared and optimized. Methods evaluated will include a variation of the immunoaffinity method. Extraction/concentration and enzyme immunoassay protocols developed will be evaluated using actual seafood and environmental sample.